Immunostimulatory combinations and methods

Abstract

The invention provides immunostimulatory compositions and methods of enhancing TLR8-mediated biological activity. Generally, the immunostimulatory compositions include a TLR8 agonist and an immunostimulatory oligonucleotide in an amount effective to enhance TLR8-mediated biological activity. The invention also provides methods of inducing TLR8-mediated biological activity in immune cells. Generally, the methods include contacting immune cells with an immunostimulatory composition that includes a TLR8 agonist and an immunostimulatory oligonucleotide in an amount effective to enhance TLR8-mediated biological activity. In some cases, the immunostimulatory composition provides a synergistic enhancement of TLR8-mediated biological activity.

1 . An immunostimulatory combination that comprises:
 a TLR8 agonist in an amount that, in combination with an immunostimulatory oligonucleotide, is capable of inducing at least one TLR8-mediated biological activity; and   an immunostimulatory oligonucleotide in an amount effective to increase the extent to which the TLR8 agonist induces the at least one TLR8-mediated biological activity;   wherein the TLR8 agonist comprises a substituted imidazoquinoline amine, a tetrahydroimidazoquinoline amine, an imidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a 6,7-fused cycloalkylimidazopyridine amine, an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, a thiazolonaphthyridine amine, a pyrazolopyridine amine, a pyrazoloquinoline amine, a tetrahydropyrazoloquinoline amine, a pyrazolonaphthyridine amine, or a tetrahydropyrazolonaphthyridine amine.   
     
     
         2 . The immunostimulatory combination of  claim 1  wherein the immunostimulatory oligonucleotide comprises a CpG oligodinucleotide. 
     
     
         3 . The immunostimulatory combination of  claim 2  wherein the CpG oligodinucleotide comprises a CpG-A oligodinucleotide. 
     
     
         4 . The immunostimulatory combination of  claim 2  wherein the CpG oligodinucleotide comprises a CpG-B oligodinucleotide. 
     
     
         5 . The immunostimulatory combination of  claim 2  wherein the CpG oligodinucleotide comprises a CpG-C oligodinucleotide. 
     
     
         6 . The immunostimulatory combination of  claim 1  wherein the immunostimulatory oligonucleotide comprises a poly(T) oligonucleotide, a poly(A) oligonucleotide, or poly(C) oligonucleotide. 
     
     
         7 . The immunostimulatory combination of  claim 1  wherein the TLR8 agonist and the immunostimulatory oligonucleotide are provided in a single formulation. 
     
     
         8 . An immunostimulatory combination that comprises:
 a TLR8 agonist in an amount that, in combination with an immunostimulatory oligonucleotide, is capable of inducing at least one TLR8-mediated biological activity; and   an immunostimulatory oligonucleotide in an amount effective to increase the extent to which the TLR8 agonist induces the at least one TLR8-mediated biological activity;   wherein the immunostimulatory oligonucleotide comprises an oligonucleotide that is other than a CpG oligonucleotide.   
     
     
         9 . The immunostimulatory combination of  claim 8  wherein the TLR8 agonist comprises an imidazoquinoline amine, a tetrahydroimidazoquinoline amine, an imidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a 6,7-fused cycloalkylimidazopyridine amine, an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, a thiazolonaphthyridine amine, a pyrazolopyridine amine, a pyrazoloquinoline amine, a tetrahydropyrazoloquinoline amine, a pyrazolonaphthyridine amine, or a tetrahydropyrazolonaphthyridine amine. 
     
     
         10 . The immunostimulatory combination of  claim 9  wherein the TLR8 agonist comprises an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, or a thiazolonaphthyridine amine. 
     
     
         11 . The immunostimulatory combination of  claim 9  wherein the TLR8 agonist comprises a thiazoloquinoline amine. 
     
     
         12 . The immunostimulatory combination of  claim 8  wherein the TLR8 agonist and the immunostimulatory oligonucleotide are provided in a single formulation. 
     
     
         13 . An immunostimulatory combination that comprises:
 a TLR8 agonist in an amount that, in combination with an immunostimulatory oligonucleotide, is capable of inducing at least one TLR8-mediated biological activity; and   an immunostimulatory oligonucleotide in an amount effective to provide a synergistic increase in the at least one TLR8-mediated biological activity induced by the TLR8 agonist.   
     
     
         14 . The immunostimulatory combination of  claim 13  wherein the synergistic increase in TLR8-mediated biological activity is at least two-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         15 . The immunostimulatory combination of  claim 13  wherein the synergistic increase in TLR8-mediated biological activity is at least three-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         16 . The immunostimulatory combination of  claim 13  wherein the synergistic increase in TLR8-mediated biological activity is at least five-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         17 . A method of inducing TLR8-mediated biological activity in immune cells, the method comprising:
 contacting the immune cells with an immunostimulatory combination that comprises a TLR8 agonist and an immunostimulatory oligonucleotide in an amount effective to increase a TLR8-mediated biological activity of the cells to a greater extent than contacting the immune cells with the TLR8 agonist without the immunostimulatory oligonucleotide;   wherein the TLR8 agonist comprises an IRM compound that comprises a substituted imidazoquinoline amine, a tetrahydroimidazoquinoline amine, an imidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a 6,7-fused cycloalkylimidazopyridine amine, an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, or a thiazolonaphthyridine amine.   
     
     
         18 . The method of  claim 17  wherein the immunostimulatory oligonucleotide comprises a CpG oligodinucleotide. 
     
     
         19 . The method of  claim 18  wherein the CpG oligodinucleotide comprises a CpG-A. 
     
     
         20 . The method of  claim 18  wherein the CpG oligodinucleotide comprises a CpG-B oligodinucleotide. 
     
     
         21 . The method of  claim 18  wherein the CpG oligodinucleotide comprises a CpG-C oligodinucleotide. 
     
     
         22 . The method of  claim 17  wherein the immune cells comprise PBMCs or monocyte-derived dendritic cells. 
     
     
         23 . The method of  claim 17  wherein the TLR8-mediated biological activity comprises synthesis of a cytokine, synthesis of a chemokine, synthesis of co-stimulatory markers, maturation of antigen-presenting cells, or proliferation of B lymphocytes. 
     
     
         24 . The method of  claim 23  wherein the cytokine comprises TNF or IL-12. 
     
     
         25 . The method of  claim 17  wherein contacting the immune cells with an immunostimulatory combination comprises adding the immunostimulatory combination to isolated immune cells in vitro. 
     
     
         26 . The method of  claim 17  wherein contacting the immune cells with an immunostimulatory combination comprises administering the immunostimulatory combination to a subject in a manner that permits the immunostimulatory combination to contact immune cells of the subject in vivo. 
     
     
         27 . The method of  claim 17  wherein the immunostimulatory oligonucleotide comprises a poly(T) oligonucleotide, a poly(A) oligonucleotide, or a poly(C) oligonucleotide. 
     
     
         28 . The method of  claim 17  wherein the IRM compound and the immunostimulatory oligonucleotide form an IRM-immunostimulatory oligonucleotide complex. 
     
     
         29 . The method of  claim 28  wherein the IRM-immunostimulatory oligonucleotide complex includes intercalation of the IRM compound into the immunostimulatory oligonucleotide. 
     
     
         30 . A method of inducing TLR8-mediated biological activity in immune cells, the method comprising:
 contacting the immune cells with an immunostimulatory combination that comprises a TLR8 agonist and an immunostimulatory oligonucleotide in an amount effective to increase a TLR8-mediated biological activity of the cells to a greater extent than contacting the immune cells with the TLR8 agonist without the immunostimulatory oligonucleotide;   wherein the immunostimulatory oligonucleotide comprises an oligonucleotide other than a CpG oligonucleotide.   
     
     
         31 . The method of  claim 30  wherein the TLR8 agonist comprises an imidazoquinoline amine, a tetrahydroimidazoquinoline amine, an imidazopyridine amine, a 1,2-bridged imidazoquinoline amine, a 6,7-fused cycloalkylimidazopyridine amine, an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, a thiazolonaphthyridine amine, a pyrazolopyridine amine, a pyrazoloquinoline amine, a tetrahydropyrazoloquinoline amine, a pyrazolonaphthyridine amine, or a tetrahydropyrazolonaphthyridine amine. 
     
     
         32 . The method of  claim 31  wherein the TLR8 agonist comprises an imidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amine, an oxazoloquinoline amine, a thiazoloquinoline amine, an oxazolopyridine amine, a thiazolopyridine amine, an oxazolonaphthyridine amine, or a thiazolonaphthyridine amine. 
     
     
         33 . The method of  claim 31  wherein the TLR8 agonist comprises a thiazoloquinoline amine. 
     
     
         34 . The method of  claim 30  wherein the TLR8 agonist and the immunostimulatory oligonucleotide form a complex in which the TLR8 agonist intercalates into the immunostimulatory oligonucleotide. 
     
     
         35 . A method of inducing TLR8-mediated biological activity in immune cells, the method comprising:
 contacting the immune cells with an immunostimulatory combination that comprises a TLR8 agonist and an immunostimulatory oligonucleotide in an amount effective to provide a synergistic increase in the at least one TLR8-mediated biological activity induced by the TLR8 agonist.   
     
     
         36 . The method of  claim 35  wherein the synergistic increase in TLR8-mediated biological activity is at least two-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         37 . The method of  claim 35  wherein the synergistic increase in TLR8-mediated biological activity is at least three-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         38 . The method of  claim 35  wherein the synergistic increase in TLR8-mediated biological activity is at least five-fold over that induced by the TLR8 agonist without the immunostimulatory oligonucleotide. 
     
     
         39 . The method of  claim 36  wherein the TLR8 agonist and the immunostimulatory oligonucleotide form a complex in which the TLR8 agonist intercalates into the immunostimulatory oligonucleotide.